Flow cytometry is a scientific tool that is widely utilized in clinical and research laboratories to enable the phenotypic and functional characterization of cells by identifying specific molecules (proteins, DNA and RNA) expressed on the surface of the cell, or in the cytoplasm and even the nucleus. Fluorescence-based flow cytometry was developed in the late 1960s and first utilized as a research tool in 1970 by Leonard Herzenberg at Stanford University. Since that time, this technology has evolved rapidly revolutionizing our understanding of immunology, hematology and cell biology. Beginning with three parameter analyses in the 1970’s, the latest instruments can capture information on over 40 parameters per cell within a few seconds to minutes using as little as 100-500μL of each sample. In the clinical laboratory, these advances have been successfully leveraged to aid in the rapid diagnosis of complex immunologic disorders such as primary immunodeficiency diseases. As the resolution capacity of the flow cytometers has advanced, so has the complexity involved in data acquisition and analyses. Assay set up (which is predominantly manual in nature), sample acquisition on the flow cytometer and subsequent analyses and interpretation of complex data sets can take time and requires specialized training and expertise.
Learning Objectives:
- Outline the basic principles of flow cytometry
- Demonstrate how flow cytometry is used to define the normal and aberrant phenotypic properties of cells (immunophenotyping)
- Elucidate the power of flow cytometry to help characterize the normal and aberrant signaling properties of cells (functional analyses)
